PMID: 3772107Oct 23, 1986Paper

HBsAg: anti-HBs immune complexes. A method for separating the constituent components and assessment of the affinity of the antibody

Journal of Immunological Methods
D N HogbenM W Steward

Abstract

A number of chemical disruption agents were assessed for their ability to dissociate HBsAg:anti-HBs immune complexes and to release both the antibody and antigen component in immunologically active forms. The most appropriate reagent was 0.1 M diethylamine which could elute up to 81% of anti-HBs antibody bound to solid-phase HBsAg and retained 93% of its antigen-combining activity. Complexes formed at various degrees of antigen excess and pre-exposed to 0.1 M diethylamine at room temperature for 18 h before ultracentrifugation on sucrose density gradients were effectively dissociated. The released antibody and antigen banded at their expected densities. However, the affinity of the isolated antibody for the detergent-solubilized polypeptide complex from purified HBsAg (gp30/p25) and cyclical peptides representing amino acids 124-137 and 139-147 of HBsAg were found to be considerably lower than that of the original pooled anti-HBs immunoglobulin used to form the immune complexes. These results suggest that the highest affinity antibody subpopulation may not be completely dissociated from the complex. Care should thus be exercised in the interpretation of the significance of the observed affinity of the antibody isolated by this ...Continue Reading

References

Jan 1, 1979·Journal of Immunological Methods·J Benveniste, C Bruneau
Jan 1, 1979·Journal of Immunological Methods·W Pernice, H H Sedlacek
Sep 1, 1979·The Journal of General Virology·J SkellyA J Zuckerman
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Dec 1, 1973·Journal of Immunological Methods·S GazeM W Steward
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Jan 1, 1980·Journal of Immunological Methods·B W MaidmentT M Chu

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Citations

Oct 23, 1987·Journal of Immunological Methods·H Haas, M Schlaak
Dec 3, 1993·Journal of Immunological Methods·D GoldblattM W Turner
May 1, 1992·European Journal of Clinical Investigation·M RadkowskiJ Slusarczyk
Jan 1, 1989·Critical Reviews in Clinical Laboratory Sciences·T M Phillips
Aug 1, 1990·Clinical and Experimental Immunology·S Hunt GerardoR H Stevens

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